聚酰胺色谱法分离油茶蒲提取物中抑制5α-还原酶的活性部位

Separation of 5α-Reductase Inhibitor from Oiltea Camellia Extracts by Polyamide Chromatography

采用聚酰胺色谱法研究油茶蒲提取物(oiltea camellia extracts,OCE)中抑制5α-还原酶的活性部位的分离方法。以体积比为3∶2∶2∶1的正丁醇-甲醇-水-冰醋酸为展开剂,0.5 g/100 mL FeCl3乙醇溶液为显色剂,采用聚酰胺色谱法将OCE清晰地分成Rf为0.85~0.037 5的7个分离部位Fr1~Fr7。5α-还原酶抑制活性的检测结果显示,Fr4和Fr5抑制活性最强,抑制率分别为(73.01±2.93)%和(76.28±1.37)%,较OCE纯化前的抑制率(57.79±1.67)%显著增加,其中Fr5得率为2.10%,且HPLC图谱中峰型较为单一,活性成分得到富集。结果表明,聚酰胺色谱法能够有效分离OCE中抑制5α-还原酶的活性部位,可为进一步制备OCE中抑制5α-还原酶的关键化合物奠定基础,同时也为强极性酚酸类混合物体系的分离提供了一种简便、高效的方法。

This study reports the separation of fractions with inhibitory activity against 5α-reductase from oiltea camellia extracts(OCE), prepared by refluxing the fruit hull of oiltea camellia(Camellia oleifera Abel.) with hot ethanol. After polyamide chromatography and color development using n-butyl alcohol-methanol-water-glacial acetic acid volume ratio 3:2:2:1, OCE were clearly divided into seven fractions, namely Fr1 through Fr7, whose Rf was in t he range of 0.85–0.037 5. The results of inhibition assays indicated that Fr 4 and Fr 5 had the highest inhibitory effect on 5α-reductase, with inhibition rates of(73.01 ± 2.93)% and(76.28 ± 1.37)%, respectively, which were signifi cantly higher than that reported for OCE(57.79 ± 1.67)%. The yield of Fr5 was 2.10% and its composition wa s relatively simple by HPLC detection, suggesting successful concentration of active ingredients. These results suggest that the application of polyamide chromatography can effectively separate anti-5α-reductase components from OCE, and provides a basic condition for the preparation of key anti-5α-reductase compounds from OCE and a convenient and effi cient method of separating strongly polar phenolic acids system as well.